Ultrasensitive and Multiplexed Target Detection Strategy Based on Photocleavable Mass Tags and Mass Signal Amplification
Co-infections pose significant challenges not only clinically, but also in terms of simultaneous diagnoses. The development of sensitive, multiplexed analytical platforms is critical for accurately detecting viral co-infections, particularly in complex biological environments. In this study, we pres...
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MDPI AG
2025-07-01
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| Series: | Nanomaterials |
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| author | Seokhwan Ji Jin-Gyu Na Woon-Seok Yeo |
| author_facet | Seokhwan Ji Jin-Gyu Na Woon-Seok Yeo |
| author_sort | Seokhwan Ji |
| collection | DOAJ |
| description | Co-infections pose significant challenges not only clinically, but also in terms of simultaneous diagnoses. The development of sensitive, multiplexed analytical platforms is critical for accurately detecting viral co-infections, particularly in complex biological environments. In this study, we present a mass spectrometry (MS)-based detection strategy employing a target-triggered hybridization chain reaction (HCR) to amplify signals and in situ photocleavable mass tags (PMTs) for the simultaneous detection of multiple targets. Hairpin DNAs modified with PMTs and immobilized loop structures on magnetic particles (Loop@MPs) were engineered for each target, and their hybridization and amplification efficiency was validated using native polyacrylamide gel electrophoresis (PAGE) and laser desorption/ionization MS (LDI-MS), with silica@gold core–shell hybrid (SiAu) nanoparticles being employed as an internal standard to ensure quantitative reliability. The system exhibited excellent sensitivity, with a detection limit of 415.12 amol for the hepatitis B virus (HBV) target and a dynamic range spanning from 1 fmol to 100 pmol. Quantitative analysis in fetal bovine serum confirmed high accuracy and precision, even under low-abundance conditions. Moreover, the system successfully and simultaneously detected multiple targets, i.e., HBV, human immunodeficiency virus (HIV), and hepatitis C virus (HCV), mixed in various ratios, demonstrating clear PMT signals for each. These findings establish our approach as a robust and reliable platform for ultrasensitive multiplexed detection, with strong potential for clinical and biomedical research. |
| format | Article |
| id | doaj-art-4682e00453314fc58231fbff10305fea |
| institution | Kabale University |
| issn | 2079-4991 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | MDPI AG |
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| series | Nanomaterials |
| spelling | doaj-art-4682e00453314fc58231fbff10305fea2025-08-20T03:36:27ZengMDPI AGNanomaterials2079-49912025-07-011515117010.3390/nano15151170Ultrasensitive and Multiplexed Target Detection Strategy Based on Photocleavable Mass Tags and Mass Signal AmplificationSeokhwan Ji0Jin-Gyu Na1Woon-Seok Yeo2Department of Bioscience and Biotechnology, Bio/Molecular Informatics Center, Konkuk University, Seoul 05029, Republic of KoreaDepartment of Bioscience and Biotechnology, Bio/Molecular Informatics Center, Konkuk University, Seoul 05029, Republic of KoreaDepartment of Bioscience and Biotechnology, Bio/Molecular Informatics Center, Konkuk University, Seoul 05029, Republic of KoreaCo-infections pose significant challenges not only clinically, but also in terms of simultaneous diagnoses. The development of sensitive, multiplexed analytical platforms is critical for accurately detecting viral co-infections, particularly in complex biological environments. In this study, we present a mass spectrometry (MS)-based detection strategy employing a target-triggered hybridization chain reaction (HCR) to amplify signals and in situ photocleavable mass tags (PMTs) for the simultaneous detection of multiple targets. Hairpin DNAs modified with PMTs and immobilized loop structures on magnetic particles (Loop@MPs) were engineered for each target, and their hybridization and amplification efficiency was validated using native polyacrylamide gel electrophoresis (PAGE) and laser desorption/ionization MS (LDI-MS), with silica@gold core–shell hybrid (SiAu) nanoparticles being employed as an internal standard to ensure quantitative reliability. The system exhibited excellent sensitivity, with a detection limit of 415.12 amol for the hepatitis B virus (HBV) target and a dynamic range spanning from 1 fmol to 100 pmol. Quantitative analysis in fetal bovine serum confirmed high accuracy and precision, even under low-abundance conditions. Moreover, the system successfully and simultaneously detected multiple targets, i.e., HBV, human immunodeficiency virus (HIV), and hepatitis C virus (HCV), mixed in various ratios, demonstrating clear PMT signals for each. These findings establish our approach as a robust and reliable platform for ultrasensitive multiplexed detection, with strong potential for clinical and biomedical research.https://www.mdpi.com/2079-4991/15/15/1170hybridization chain reactionmass spectrometrymass tagsmultiplexed detectionphotocleavablesignal amplification |
| spellingShingle | Seokhwan Ji Jin-Gyu Na Woon-Seok Yeo Ultrasensitive and Multiplexed Target Detection Strategy Based on Photocleavable Mass Tags and Mass Signal Amplification Nanomaterials hybridization chain reaction mass spectrometry mass tags multiplexed detection photocleavable signal amplification |
| title | Ultrasensitive and Multiplexed Target Detection Strategy Based on Photocleavable Mass Tags and Mass Signal Amplification |
| title_full | Ultrasensitive and Multiplexed Target Detection Strategy Based on Photocleavable Mass Tags and Mass Signal Amplification |
| title_fullStr | Ultrasensitive and Multiplexed Target Detection Strategy Based on Photocleavable Mass Tags and Mass Signal Amplification |
| title_full_unstemmed | Ultrasensitive and Multiplexed Target Detection Strategy Based on Photocleavable Mass Tags and Mass Signal Amplification |
| title_short | Ultrasensitive and Multiplexed Target Detection Strategy Based on Photocleavable Mass Tags and Mass Signal Amplification |
| title_sort | ultrasensitive and multiplexed target detection strategy based on photocleavable mass tags and mass signal amplification |
| topic | hybridization chain reaction mass spectrometry mass tags multiplexed detection photocleavable signal amplification |
| url | https://www.mdpi.com/2079-4991/15/15/1170 |
| work_keys_str_mv | AT seokhwanji ultrasensitiveandmultiplexedtargetdetectionstrategybasedonphotocleavablemasstagsandmasssignalamplification AT jingyuna ultrasensitiveandmultiplexedtargetdetectionstrategybasedonphotocleavablemasstagsandmasssignalamplification AT woonseokyeo ultrasensitiveandmultiplexedtargetdetectionstrategybasedonphotocleavablemasstagsandmasssignalamplification |